Fig. 8. MOM recruitment assay with FL-3/7, p120(ctn) and MOM-N-cadherin. MCF7 cells were co-transfected with pMOM-N-cadherin, mARVCF splice variant FL-3/7 fused to an Xpress-tag and p120(ctn) as a EGFP-fusion protein. Cells were stained with a Xpress antibody to detect FL-3/7. As shown in B, FL-3/7-Xpress (a) and EGFP-p120(ctn) (b) can bind to MOM-N-cadherin (not stained) in the same cell. In c, the merged image of a and b is shown. (A) Cells were treated as in B and pictures were taken where both FL-3/7-Xpress (a) and EGFP-p120(ctn) (b) were detected at the plasma membrane. (c) The merged image of (a) and (b). To confirm that these cells were also positive for MOM-N-cadherin, the same coverslips were washed again and stained for MOM-N-cadherin with the birch profilin antibody (BP); the positive BP staining is given in d. (e) The merged image of a, b and d where the MOM-N-cadherin staining is coloured artificially blue. In f, 500 cells expressing FL-3/7-Xpress, EGFP-p120(ctn) and MOM-N-cadherin were counted. In 32.9% (mARVCF) or 35.2% (p120(ctn)) of these cells a mitochondrial located interaction of both constructs was detected. In 67.1% (mARVCF) or 64.8% (p120(ctn)) of cells expressing MOM-N-cadherin, EGFP-FL-3/7 and EGFP-p120(ctn) such an interaction did not take place. Error bars indicate s.d. Bar, 10 µm.
